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弓形虫特异性多克隆鸡卵黄免疫球蛋白的生产、特性及应用。

Production, characterization and applications for Toxoplasma gondii-specific polyclonal chicken egg yolk immunoglobulins.

机构信息

Laboratory of Immunoparasitology, Institute of Biomedical Sciences, Universidade Federal de Uberlândia, Minas Gerais, Brazil.

出版信息

PLoS One. 2012;7(7):e40391. doi: 10.1371/journal.pone.0040391. Epub 2012 Jul 12.

Abstract

BACKGROUND

Toxoplasma gondii may cause abortions, ocular and neurological disorders in warm-blood hosts. Immunized mammals are a wide source of hyperimmune sera used in different approaches, including diagnosis and the study of host-parasite interactions. Unfortunately, mammalian antibodies present limitations for its production, such as the necessity for animal bleeding, low yield, interference with rheumatoid factor, complement activation and affinity to Fc mammalian receptors. IgY antibodies avoid those limitations; therefore they could be an alternative to be applied in T. gondii model.

METHODOLOGY/PRINCIPAL FINDINGS: In this study we immunized hens with soluble tachyzoite antigens of T. gondii (STAg) and purified egg yolk antibodies (IgY) by an inexpensive and simple method, with high yield and purity degree. IgY anti-STAg antibodies presented high avidity and were able to recognize a broad range of parasite antigens, although some marked differences were observed in reactivity profile between antibodies produced in immunized hens and mice. Interestingly, IgY antibodies against Neospora caninum and Eimeria spp. did not react to STAg. We also show that IgY antibodies were suitable to detect T. gondii forms in paraffin-embedded sections and culture cell monolayers.

CONCLUSIONS/SIGNIFICANCE: Due to its cost-effectiveness, high production yield and varied range of possible applications, polyclonal IgY antibodies are useful tools for studies involving T. gondii.

摘要

背景

刚地弓形虫可能会导致温血宿主流产、眼部和神经系统疾病。免疫哺乳动物是大量高免血清的来源,这些血清可用于多种方法,包括诊断和宿主-寄生虫相互作用的研究。不幸的是,哺乳动物抗体在生产方面存在一些局限性,例如需要动物采血、产量低、与类风湿因子、补体激活和 Fc 哺乳动物受体的亲和力干扰等。IgY 抗体避免了这些局限性;因此,它们可能是替代方法,可应用于弓形虫模型。

方法/主要发现:在这项研究中,我们用廉价且简单的方法,用高产量和高纯度对刚地弓形虫可溶性速殖子抗原(STAg)和纯化的蛋黄抗体(IgY)进行免疫母鸡。IgY 抗-STAg 抗体具有高亲和力,并能识别广泛的寄生虫抗原,尽管在免疫母鸡和小鼠产生的抗体的反应谱上观察到一些明显的差异。有趣的是,针对新生隐球菌和艾美耳球虫的 IgY 抗体不与 STAg 反应。我们还表明,IgY 抗体适用于检测石蜡包埋切片和培养细胞单层中的弓形虫。

结论/意义:由于其成本效益高、产量高和可能的应用范围广泛,多克隆 IgY 抗体是研究弓形虫的有用工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/65fb/3395712/2e328d002e24/pone.0040391.g001.jpg

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